The current methods of isolation include sourcing stem cells from the following; adult bone marrow, adipose tissue, fibroblastic tissues, blood, embryonic tissue. Adult differentiated cells are artificially induced to become more embryonic-like by transferring genetic material into the cells such as, for example, induced pluripotent stem cells (iPSCs). Non-embryonic stem cells are artificially induced to differentiate into the desired tissue by transferring genetic material into these cells. To prevent teratoma formation embryonic stem cells and induced-pluripotent stem cells are encouraged to differentiate using inductive agents either singly or in combination. Adult stem cells are also encouraged to differentiate using inductive agents, either singly or in combination. Induced embryonic, induced pluripotent, and adult stem cells are propagated and then introduced into the damaged tissue, which they then replace. However, such current methods of isolation and introduction of embryonic, iPSC, and adult stem cells have multiple inherent problems.
Embryonic sourcing of stem cells is highly controversial and subject to strict regulation concerning their collection and use. Use of adult derived stem cells engender less controversy and scrutiny, but current methods of obtaining and using them still presents several problems.
Obtaining stem cells from adult differentiated cells require using viruses to transfer embryonic genes into the differentiated cells. These artificially created cells, for example, iPSCs, can form teratomas when placed into adult tissues in an undifferentiated state. In addition, the viruses used for transfer of the genetic material can go “rogue” causing multiple problems within the altered cells. Instances involving transfer of genetic material by viruses for tissue repair are rife with problems such as infections and cancer.
The current invention utilizing mesodermal stem cells (MesoSCs) and nucleated plasma particles (Nuc-P2s) derived from whole blood, circumvents the dangers inherent in using viruses and foreign genetic material. Neither adult MesoSCs nor Nuc-P2s manipulate the genetics of the damaged tissues, but incorporate into the recipient's own healthy tissue where they are introduced.
Currently, the principal methods of obtaining adult stem cells include bone marrow aspiration, liposuction from adipose tissue, enzymatic digestion of solid tissue biopsies, and isolation from blood. The first three methods of adult stem cell collection are problematic because they are invasive and require the use of anesthesia and surgical expertise. They also increase the risk the patient may experience complications and pain from the extraction procedure. Enzymatic digestion of biopsy tissue may leave remnants of tissue proteins and/or enzymes, which may elicit an immune response.
While less expensive, time consuming and invasive, collection of stem cells by the use of venipuncture causes an immediate clotting response, reducing the number of adult stem cells in the sample. Consequently, while existing stem cell isolation from blood is far less expensive and less invasive than other methods currently in use, the yield of stem cells is much lower.
The existing methods of treating damaged tissue with stem cells suffer from problems similar to the problems of stem cell isolation methods. Treatment of damaged central nervous system tissue by stem cells is currently done by a craniotomy, intra-thecal transplant or intra-nasally. A craniotomy adds the risks and expense of neurosurgery. In intra-thecal transplants, the exact volume of cerebral spinal fluid must be returned to the patient or they risk an extended period of severe headaches. Current intra-nasal application requires the use of hyaluronidase or mannitol, which compromises the function of nasal tissue, and risks the patient contracting meningitis. Intravenous introduction of stem cells derived from adipose tissue have the potential to cause death due to a fat embolism and intravenous introduction of stem cells cannot repair damage to the brain and spinal cord due to an inability to pass through the blood-brain barrier. Intravenous introduction of stem cells cannot repair damage to the epithelia lining the lung alveoli due to the gas/liquid basement membrane barrier between the capillaries and alveoli. Furthermore, during intravenous injection of stem cells during treatment, the clotting response reduces the number of adult stem cells reaching the treatment area. Currently, few advances have been made in optimizing the cell's reparative activities and proliferation rates to increase the number of effective stem cells used to treat damaged tissues.
Accordingly, what is needed are alternative methods which increase the numbers and availability of adult MesoSCs and Nuc-P2s in the blood circulation for use in repairing tissue damage via collected MesoSCs and Nuc-P2s separated by size and suitability for specific types of tissue repair, and optimizing the availability of delivered MesoSCs and/or nucleated plasma particles to damaged tissue. The present invention addresses these needs by increasing the available levels of adult MesoSCs and Nuc-P2s and optimizing the conditions for collecting the same. Further, the present invention allows for isolation of the different types of nucleated plasma particles and MesoSCs and their targeted introduction into damaged tissues in a manner that maximizes regeneration of damaged tissues.